Malonyl derivatives of 6-({60 -aminoacylamido)penicillanic acids

ABSTRACT

The malonyl derivatives of 6-( Alpha aminoacylamido)penicillanic acids, the non-toxic salts and esters thereof, are useful as antibacterial agents, therapeutic agents in animals including man, of particular value against gramnegative bacteria and as animal feed nutritional supplements.

United States Patent [1 1 Rosati MALONYL DERIVATIVES OF 6-(a-AMINOACYLAMIDO)PENICILLANIC ACIDS [75] Inventor: Robert L. Rosati, Stonington, Conn.

[73] Assignee: Pfizer Inc., New York, NY.

[22] Filed: Feb. 8, 1973 [21] Appl. No.: 330,751

52 US. Cl 260/239.1; 424/271; 260/243 0 51 Int. cm c070 499/44 [58] Field of Search 260/239.1

[56] References Cited UNITED STATES PATENTS 3,483,188 12/1969 McGregor ..260/239.1

[ Dec. 23, 1975 Primary ExaminerNich0las S. Rizzo Attorney, Agent, or Firm-Connolly and Hutz [57] T ABSTRACT 3 Claims, No Drawings MALONYL DERIVATIVES OF 6-(a-AMINOACYLAMIDO)PENICILLANIC ACIDS BACKGROUND OF THE INVENTION A large number of 6-(a-aminoacylamido)penicillanic acids wherein the acyl moiety is alkanoyl or substituted alkanoyl wherein the substituent is an aryl, cycloalkyl or heterocyclic group are disclosed in U.S. Pat. Nos. 2,985,648; 3,007,920; 3,192,198; 3,485,819; 3,342,677; 3,538,083; 3,553,201; British Pat. Nos. 873,049; 903,785; 991,586; 1,033,257 and 1,189,990. Further, 6-[a-substituted amino)acylamido]penicillanic acids are described in U.S. Pat. Nos. 3,198,788; 3,248,387; 3,308,023; 3,325,477; 3,340,252; 3,381,001; 3,433,784; 3,518,253; British Pat. Nos. 891,977; 894,457; 985,688; 1,048,907; 1,051,675; 1,057,697; 1,064,893; 1,066,107; 1,125,339; 1,180,745; 1,210,472; Belgian Pat. No. 593,295 and Japanese Pat. No. 7,] 16,994. Additionally, 6-(a-ureido acylamido)penicillanic acids are disclosed in U.S. Pat. No. 3,352,857 and German Pat. No. 2,054,772; 6-(aguanidinoacylamido)penicillanic acids in U.S. Pat. Nos. 3,454,557 and 3,406,185; and a variety of pquanidinoaroyl-, p-guanidinomethylarolyand pguanidinoarylalkanoylamido-penicillanic acids are disclosed in U.S. Pat. No. 3,543,265. British Pat. No. l ,061 ,335 discloses 6-( D-a-hydrazinocarbonylaminoaphenylacetamido)penicillanic acid, and British Pat.

No. 1,053,818 describes esters of 6-(01- oxalamidoacylamido)penicillanic acids.

A wide variety of 6- a-( 3-substituted ureido)acylamidolpenicillanic acids and 6-[a-(3-substituted thioureido)acylamido]penicil1anic acids are reported in the recent literature. U.S. Pat. Nos. 3,479,339; 3,481,922; Netherlands Pat. Nos. 6,901,646; 6,908,909; and Japanese Pat. No. 7,112,732 describe such compounds wherein the 3- substituent is a carbamoyl group; the compounds being referred to as 6-[a-(3-allophanamido)acylamid'o]- penicillanic acids. U.S. Pat. No. 3,579,501 discloses 6-[a-(3 -guanylureido)acylamido]penicillanic acids; that is, such compounds wherein 3-substituent is a guanyl moiety.

The above described products are active as antibacterial agents against a variety of gram-negative bacteria. However, they are more active in vivo via the intraperitoneal route of administration than the oral route.

SUMMARY OF THE INVENTION There has now been found a novel series of antibacterial agents; namely, malonyl derivatives of 6-(aaminoacylamide)penicillanic acids of the formula wherein R is selected from the group consisting of hydrogen and acyloxy lower alkyl wherein acyloxy is selected from the group consisting of lower a1- kanoyloxy, benzoyloxy and substituted benzoyloxy wherein the substituent is selected from the group consisting of chloro, bromo, fluoro, lower alkyl, lower alkoxy and trifluoromethyl; cations selected from the group consisting of sodium, potassium, calcium, magnesium and ammonium; and amines selected from the group consisting or procaine, N,N'-dibenzylethylenediamine, N-ethylpiperidine, dibenzylamine, N,Nbis(dehydroabietyl)ethylenediamine, l-ephenamine, triethylamine and benzhydrylamine.

R is selected from the group consisting of hydrogen, alkyl of l to 14 carbon atoms, cycloalkyl of 3 to 6 carbon atoms, 1,4-cyclohexadienyl, naphthyl, cyclohepatrienyl, benzyl, phenethyl, indolymethyl, furyl, thienyl, w-ethylthio( lower alkyl and wherein Y is selected from the group consisting of hydrogen, nitro, di(lower alky1)amino, lower a1- kanoylamine, lower alkyl, lower alkoxy, hydroxy, sulfamyl, chloro, bromo, fluoro, iodo and trifluoromethyl);

R is selected from the group consisting of hydrogen and lower alkyl;

R and R when taken together with the carbon atom to which they are attached are cycloalkylidene of 3 to 10 carbon atoms;

R is a member selected from the group consisting of hydrogen, alkyl having from 1 to 4 carbon atoms, cycloalkyl having from 3 to 6 carbon atoms, phenyl, nitrophenyl, naphthyl, pyridyl, anthranyl, furyl, thienyl, indolylmethyl, thiazolyl, pyrimidinyl, pyradizinyl, biphenyl, indazolyl and mono and disubstituted phenyl wherein each substituent is chlorine, bromine, fluorine, alkyl having from 1 to 3 carbon atoms, hydroxy, cyano, amino, trifluoromethyl, acetamido, methoxycarbonyloxy or methoxy; and v Q is a member selected from the group consisting of hydrogen, alkyl having from 1 to. 4 carbon atoms, naphthyl, quinolyl, phenyl, biphenyl, indanyl or 2-( 1,4-naphthoquinonyl); cations selected from the group consisting of sodium, potassium, calcium, magnesium and ammonium; and amines selected from the group consisting of procaine, N,N dibenzylethylenediamine, N-ethylpiperidine, dibenzylamine, N,N-bis( dehydroabietyl )ethylenediamine, l-ephenamine, triethylamine and benzhydrylamine.

Also included within the scope of this invention are the optically active isomeric forms and mixtures thereof which arise by virtue of the asymmetric a-carbon atom of the acyl side chain. These are the D- and the L-diastereoisomers and the DL-form which is a mixture of the two optically-active isomers. The D- form of these compounds is the preferred form because of its greater activity relative to that of the L-- or the DL-forms.

Other isomers of the herein described compounds in addition to those arising from the asymmetric oz-carbon of the acyl side chain are, of course, possible because of the presence of asymmetric carbon atoms'in the 6- aminopenicillanic acid nucleus.

with a carboxy group or a reactive functional derivative of a carboxy group of an appropriate malonic acid derivative of the formula wherein the variables R R R, R and Q are as defined above.

The terms lower alkyl, lower alkoxy and lower alkanoyloxy as used herein are intended to include those alkyl, alkoxy and alkanoyloxy groups having from one to four carbon atoms.

Suitable esters of the formula II reactants are those wherein R is acyloxy lower alkyl as defined above and those wherein R is a group which can readily be removed as, for example, by catalytic hydrogenation (benzyl, cyanomethyl, phenacyl, allyl and diphenylmethyl).

Suitable reactive functional derivatives of acids of formula III are the acid chlorides or bromides. The acid reactant can be reacted with a condensing agent such as a carbonylditriazole and hexahalocyclotripha triazine to give a reactive intermediate which is coupled to the 6-(a-aminoacylamido)penicillanic acid. Additionally, the appropriate acid azide or an active ester or thio ester of the formula III reactant with, for example, N-hydroxyphthalimide, N-hydroxysuccinimide, a phenol or thiophenol can be used as acylating agent.

The preferred acylation process of this invention comprises the appropriate 6-(a-aminoacylamido)- penicillanic acid compound (formula II) with the acid form of a formula II reactant in the presence of a carbodiimide for reasons of convenience, availability or reactants and overall yield or product.

The 6-(a-aminoacylamido)penicillanic acid reactant can be used in a variety of forms. It can, for example, be used as the free acid or as an alkali metal or amide salt thereof. The use of a salt form of the penicillanic acid reactant is frequently of advantage since the solubility can be manipulated by judicious choice of the salt to permit the use of aqueous or non-aqueous systems. Alkali metal salts are valuable for use in aqueous systems. In non-aqueous systems, an amine salt such as a tertiary lower alkylamine salt, e.g., thriethylamine, or

an N-alkyl piperidine salt is generally used. Alternatively, an ester of the 6-(a-aminoacylamido)penicillanic acid is used, especially in non-aqueous systems. In those instances wherein the final product (formula I) is desired in the form of an ester (R is other than hydrogen), it is obvious and practical to use that ester form of the penicillanic acid reactant.

The acylation process is conducted in a reaction inert solvent system which can be aqueous or nonaqueous. Aqueous or non-aqueous solvent systems can be used when a carbodiimide is the condensing agent. When using a carbodiimide in an aqueous system, the pH is desirably adjusted to the range of about 5 to about 8, and preferably to about 6 to 7. In a typical procedure, the formula III reactant and carbodiimide are mixed in eq'uimolar proportions in a suitable solvent (tetrahydrofuran, dioxan) and a water-miscible organic solvent solution (water plus dioxan or tetrahydrofuran) containing the 6-(a-aminoacylamido)- penicillanic acid is added at room temperature and the mixture stirred for several hours until reaction is complete. Temperatures of from about 5 to 30C. are generally used. In most instances, an excess up to about 10% of the condensing agent is used. The penicillin product is recovered by methods known to the art.

Acylation with an acid halide can also be conducted in aqueous or nonaqueous solvent systems. In aqueous systems, the reaction is generally-carried out at pH of from about 6 to about 9 and a temperature of from about 0C. to about C. It can also be performed. in unstable emulsions of water and a water-immiscible organic solvent such as methyl isobutyl ketone and lower alkyl acetates over the pH range of from about 2to about 4.

In addition to the above purely chemical techniques of acylation, a-sonochemical technique, that is, the application of vibrations of ultrasonic frequency (35,000 to 90,000 cycles per second), as described in US. Pat. No. 3,079,314, issued Feb. 26, 1963, can also be used to achieve acylation of the 6-[(oz-amino substituted) acylamido1penicillanic acid, especially acylation with an acid halide. Acylation under such conditions is rapid and permissive of a wide range of reaction media, aqueous and non-aqueous alike, homogeneous and non-homogeneous, including emulsified systems.

The acyloxy (lower alkyl) esters of formula I com pounds are prepared by the above described acylation procedures but using the appropriate acyloxy (lower alkyl) ester of the appropriate 6-(a-aminoacylamido)- penicillanic acid in place of the non-esterified 6-(aaminoacylamido)penicillanic acid. The acyloxy (lower alkyl) esters of the 6-(a-aminoacylamido)penicillanic acids are prepared according to methods described in Belgian Pat. No. 721,515 and by Daehne et al., J. Med. Chem. 13, 607-612 (1970).

The acyloxy (lower alkyl) halides are synthesized from'the corresponding acid chlorides and aldehydes or ketones in accordance with the general procedures of Ulich et al., J. Am. Chem. Soc, 43, 660 (1921) and Euranto et al., Acta. Chem. Scand. 20, I273 (I966). The formation of esters from acid salts and alkyl helides is well documented in the chemical literature (Zook and Wagner, Synthetic Organic Chemistry, John Wiley and Sons, Inc., New York, 1956, p. 484).

The substituted malonic acid reactants of the formula are prepared by the general methods described in Proc, Indian Acad. Sci. 14A, I 12-22 1941) and 22A, 4004 I945 An appropriate aldehyde is mixed with malonic acid and heated at steam bath temperature for about 4 to 6 hours. The crystalline condensation product may be recrystallized from an organic solvent such The mono-esters of these compounds are similarly prepared by condensing an appropriate aldehyde with a mono-ester of malonic acid. The mono-esters of malonic acid may be prepared from commercially availble (Ii-esters or di-esters prepared by the action of desired alcohol and a mineral acid on sodium cyanoacetate. The diester is partially saponified to yield the monoester. The mono-esters may also be prepared by react- 6 ing malonic acid with thionyl chloride to yield the mono-acid chloride which is then contacted with the desired alcohol.

In the formulae accompanying the preparation of these new penicillins, APA" represents the moiety The novel penicillins described herein exhibit in vitro activity against a wide variety of both gram-positive and gram-negative bacteria. Their useful activity can readily be demonstrated by in vitro tests against various organisms in a brain-heart infusion medium by the usual two-fold serial dilution technique. The in vitro activity of the herein-described compounds renders them useful for topical application in the form of ointments, creams and the like, or for sterilization purposes, e.g.. sick room utensils.

The in vitro spectra of a number of these compounds are presented in Table 1 below. -a-D-Aminobenzylpenicillin is included for the purpose of comparison. The compounds of Table 1 have the formula CH CO APA H.Tricthylaminc TH l TABLE I.

In Vitro Spectra of Malonamido Acyl Derivatives of a-D-Aminobenzylpenici11in Against Certain Gram-negative Organisms (MlCs) l'lculi Is. avrugi/msu I. Inirubilis R 266 490 CO 15 O 3.4dichlorophenyl 25 1.5 6 12.5 ethyl 3.4-dichlorophcnyl -5 25 5-indanyl 4-acctamidophcnyl 12.5 3.12 1.56 5'-indanyl 4-biphenyl 6.25 0.78 3.12 H.TF.A* 3.5-dimcthoxyphenyl 3.12 0.78 0.78 H.TF.A 2,6dichlorophenyl 6.25 0.39 0.78 H.TF.A Z-trifluoromethylphenyl 12.5 3.12 3. I 2 H.TEA 4fluorophcnyl 3.12 0.78 0.39 H.TF.A Z-Iluorophcnyl 12.5 0.78 1.56 H.TF.A *Lmcthylphcnyl 1.56 0.78 0.39 H.TF.A 4-mcthoxycarhonyloxyphcnyl 6.25 1.56 0.78 H.TF.A 3.4-dimethoxyphcnyl 6.25 1.56 0.78 H.TF.A 4-chlorophcnyl 3.12 1.56 0.78 H.TF.A 9-anthranyl 50 6.25 6.25 H.TF.A 3-fluorophcnyl 3. I 2 0.78 1.56 H.TF.A 4lrifluoromethylphenyl 6.25 1.56 3.12 H.TF.A 3-trillunromethylphenyl 3. I 2 0.78 1.56 H.TF.A 4-mcthylphcnyl 1.56 0.78 0.39 H.TF.A 4-cyanophcnyl 25 3.12 50 H.TF.A l-naphthyl 3.12 0.78 1.56 H.TF.A Z-naphthyl 1.56 0.78 0.78 H.TEA 3-methoxyphcnyl 6.25 1.56 0.39 H.TEA 3-mcthylphcnyl 3.12 0.78 0.39 H.TF.A 4-hromophcnyl 6.25 0.78 0.78 H.TF.A 4-nitrophcnyl 3.12 0.78 0.78 H.TF.A 4-amin0phcnyl 25 3.12 3.12 H.TF.A 3.5-dichlorophcnyl 1.56 0.39 1.56 H.TF.A

12.5 1.56 6.25 H.TF.A

-continued Route* R Organism Dosc(mg/kg) Protection Q 4-lrifluoromcthylphcnyl If. ('uli 266 200 (P) 40 H.TF.A

r; all; 266 50 (so) so I.\'. uorugl'nm'u lO4 400 (SC) ll) Ii-trifluoromclhylphcnyl If. vuli 266 200 (P0) 20 H.TF.A

I-.'. mli 266 50 (SC) 40 l.\'. uvruginmu Hi4 400 (SC) 0 P() oral subcutaneous The acute system infections in mice were produced Many of the penicillin ester compounds of this invenby the intraperitoneal inoculation of standardized culi hibit i v d absorption on oral administra- IufeS Suspended in 5% h g gastric muCiIl- Treatment 5 tion over that produced by the corresponding free acid with the drugs was initiated 0.5 hours after inoculation or lk li metal lt f Th th fore, represent of the infecting organism. A second dose was adminisv i t d effectiv dosage forms of the novel tcred four hours later. The percent protection was penicillins of formula l above. determined after a hold period of four days. Further, many of the esters, especially the acyloxy The oral and parenteral dosage levels in humans and (lower alkyl) esters described herein, although inactive an ma s for e e ein described compounds e, n or of relatively low activity against gram-negative orgeneral. on the Order Of from -200 mg/kg of body ganisms-per se are, when administered orally to'aniweight per day and from about lO-lOO mg/kg of body mals, including man, metabolized to the parent acid weight per day, respectively. For topical application, a which has a wide spectrum of activity against gramposithe dosage level is on theorder of from about 10 to 200 25 tive and gram-negative bacteria. They thus serve as mg/kg/day.

sources of the parent compounds since they are biolog- When used for the purposes described herein, the ically converted in vivo to said compounds. The rate of valuable products of this invention can be used alone or metabolic conversion of such esters to the parent acid in admixture with other antibiotics or in combination occurs at such a rate as to provide an effective and with a pharmaceutical carrier selected on the basis of prolonged concentration of the present acid in the the chosen route of administrationand standard pharanimal body. In effect, such esters act as depot sources maceutical practice. For example, they may be adminfor the parent acid. Especially useful in this respect are istered orally in the form of tablets containing such the acyloxy(lower alkyl) esters such as the benzoyloxexcipientsas starch, milk, sugar, certain types of clay, ymethyl-, acetoxymethyl-, acetoxyethyl-, pivaloyloxetc., or in capsules alone or m admixture with the same 3 5 ymethyland a-ethylbutyryloxymethyl esters.

or equivalent excipients. They may also be adminis- Alsov useful in this respect are the esters of Fonnula l tered orally in the form of elixirs or oral suspensions where Q is ethyl or 5-indanyl.

which may contain flavoring or coloring agents, or be Additionally useful compounds are analogous derivainjected parenterally, that is, for example, intramuscutives of 7-aminocephalosporanic acid, desacetoxy 7- larly or subcutaneously. For parenteral administration 40 amineeephalosporanie id, desaeety] 7- i hthey are best used in the form of a sterile solution or alosporanic acid and tertiary amine derivative f s p n h y be either aqueous Such as Water, 7aminocephalosporanic acid wherein the 3-acetoxy isotonic saline, isotonic dextrose, Ringers solution, 01' group is displaced by a tertiary amine function, as antinon-aqueous such as fatty oils of vegetable origin (cotb i agents against b h iri d ton See pe l, f", a flnd 0ther negative bacteria. They are prepared by acylation of aqueous veh cles which Wlll not'interfere with the therh appropriate 7 l(a amino substimted)acylamido]' f p efficlenc) of the Preparatlon and are non-toxic cephalosporanic acid compounds of formula IV below in the volume or propor Used (g y w i P pylene with a reactive functional derivative of an appropriate glycol, Sorbltoll- Addltlonalllf, Compositions Sultable substituted malonic acid of formula III according to for extemporaneous preparation of solutions prior to h d described h i s I R -'CH- co- NH- H- CH CH I IV NH 0: N C -CH A COOH administration may advantageously be made. Such in formula IV R is as defined above and A is selected compositions may include liquid dlluents, for example, from the group consisting of hydrogen, hydroxy; acepropylene glycol. diethyl carbonate, gylce rol, sorbitol, toxy and tertiary amino. Respresentative of the tertiary etc; buffering agents, as well as local anlsthetics and amino groups which displace the acetoxy moiety ae norga rtuc salts to afford desirable pharmacological pyridine, imidazole, benzimidazole, pyrimidine and prope |es.

substituted derivatives'of these amines and tri(alkyl)amethods for their preparation, are described in the literature;

US. Pat. Nos. 3,560,489; 3,575,969; French Pat. No. 2,032,408; J. Antibiot. Ser. A 19 (6) 243-9 (1966); Cocker et al., J. Chem. 500., 5015-5031 (1969).

EXAMPLE I 4-Chlorobenzylidene Malonic Acid p-Chlorobenzaldehyde (0.100 mole) is mixed with malonic acid (0. I mole) and heated at steam bath temperature for about 4 hours. After cooling to room temperature, the crude crystalline condensation product is recrystallized from acetonitrile, m.p. 42C. (decomp).

6-'[ D-2-Phenyl-2-( 4-Chlorobenzylidenemalonamido)Acetamido Penicillanic 1 acid-Ditriethylamine 4,-Chlorob enzylidene malonic acid (0.005 mole) in ml. of water is cooled to 0C. and the pH adjusted to 6.0 to give a clear solution. To this solution is added 6-a-aminobenzylpenicillin (0.005 mole) and 0.005 mole of l-ethyl-3-( 3-dimethylaminopropyl )-carbodiimide hydrochloride (Ott Chemical Co.). The solution is stirred for about minutes at 0C., maintaining the pH at about 6.0 with 6 N HCl. Stirring is continued for about 3 hours at room temperature, keeping the ph at about 6.0. The solution is then adjusted to pH 7.7 with saturated sodium bicarbonate solution, andextracted with several portions of ethyl acetate. The aqueous phase is overlayed with ethyl acetate and the pH adjusted to 2.7 with 6 N HCl. The separated ethyl acetate layer is washed once with water and once with saturated sodium chloride solution. To the ethyl acetate extract, dried over anhydrous sodium sulfate, is added 0.0l mole of triethylamine. The solution is taken to dryness under vacuum. The remaining ethyl acetate and triethylamine are removed from the white solid azeotropically under vacuum with several portions of methylene chloride, yield 42%.

EXAMPLE ll 4-Methoxycarbonyloxybenzylidene Malonic Acid 0.55 Mole of p-methoxycarbonyloxybenzaldehyde (Eastman) is mixed with 0.055 mole of malonic acid (Aldrich) and heated at steam bath temperature for about 4 hours. After cooling to room temperature, the crude solid is recrystallized from acetonitrile to yield yellow crystals, m.p. l94l96C. (decomp.), yield 48%.

6-[ D-2-Phenyl-2-( 4-Methoxycarbonyloxybenzylidenemalonamido)Acetamido] Penicillanic Acid'Ditriethylamine 4-Methoxycarbonyloxybenzylidene malonic acid is reacted with a-aminobenzylpenicillin by the method of Example I to yield the substituted malonyl derivative of 6-a-D-aminobenzylpenicillin, yield 48 EXAMPLE Ill 6-[D-2-Phenyl-2'( 3,5-Dichlorobenzylidenemalonamido)Acetamidol-Penicillanic acid-Ditriethylamine 3,5-Dichlorobenzylidene malonic acid 1.3 g., 0.005 mole) in 20 ml. of water is cooled to 0C, and the pH adjusted to 6.0 to give a clear solution. To this solution is added 2.0 g. (0.005 mole) of 6-a-D-aminobenzylpenicillin. After the addition of 960 mg. (0.005 mole) of 1-ethyl-3-( 3-dimethylaminopropyl )carbodiimide hydrochloride (Ott Chemical Co.), the solution is stirred for about 30 minutes at 0C., maintaining the pH at about 6.0 with 6 N HCl Stirring is continued for about 3 hours at room temperature, keeping the ph at about 6.0. The solution is thenadjusted to pH 7.7 with saturated sodium bicarbonate solution, and extracted with several portion' of ethyl acetate. The aqueous phase is overlayed with ethyl acetate and the pH adjusted 'to 2.7 with 6 N HCl The separated ethyl acetate layer is washed once with water and once with saturated sodium chloride solution. To the ethyl acetate extract, dried over sodium sulfate, is added 1.01 g. (0.0l mole)vof triethylamine. The solution is taken to dryness under vacuum. The remaining ethyl acetate and triethylamine are azeotropically removed under vacuum with 3 portions of methylenechloride. The white solid is dried under high vacuum, yield 41%.

Repetition of this procedure but replacing. 6-a- 'aminobenzylpenicillin with equimolar amounts of the appropriate acyloxyalkyl ester produces compounds of the formula @oH-co-JigA-R I NH Q-Ili EXAMPLE IV tional 4 hours at room temperature, maintaining the pH at 6.0 with 6 N HCl. The solution is then adjusted to pH 7.7 with saturated sodium bicarbonate solution, and extracted with several portions of ethyl acetate. The aqueous phase is overlayed with ethyl acetate and the pH adjusted to 2.7 with 6 N HCl. To the ethyl acetate extract, dried over sodium sulfate, is added 0.01 mole of triethylamine. The white solid remaining after removal of the solvent under vacuum is azeotropically treated with several portions of methylene chloride to remove residual moisture, yield 50%.

Repetition of this procedure but replacing 3-trifluoromethylbenzylidene malonic acid with 4-trifluoromethylbenzylidene malonic acid and Z-trifluoromcthylbenzylidene malonic acid yields the corresponding derivatives of o-a-animobenzylpenicillin.

EXAMPLE V The method of Example 1 is repeated using the appropriate a-animoacylpenicillin and the appropriate malonic acid derivative of the formula to provide a penicillin of the formula 3,4-dichlorophcnyl phcnyl 3-hromophcnyl 4aminophenyl 3-methoxy-4-hydroxyphcnyl 4-fluorophcnyl 4-methoxycarbonyloxyphcnyl 3-fluorophenyl Z-lluorophenyl 3.4dimcthoxyphcnyl 4-chlorophcnyl ."l-chlorophcnyl Z-chlorophcnyl 4-hydroxyphenyl 3,5-dimethoxyphcnyl 4-nitrophenyl 4-hiphcnyl carboxamidophcnyl indolylmethyl hydrogen Z-nitrophcnyl 4-cyanophcnyl 3-cyanophenyl lcyanophcnyl 4-hromophcnyl Z-hromophcnyl 4-acctamidophcnyl 4mcthylphcnyl .Lmclhylphcnyl Z-mcthylphcnyl l-naphthal \'l Z-naphthalyl )-anthran \'l Z-thicnyl cyclopropyl cyclohcxyl caca L F IIIIIIIIIIIIIIIIIIIIIIIIIIIIIII II IIIII Continued f: R, LI" CO APA H TH i COOH R'-C|=C/ Rl R R n s H 'l-thiazolyl u s H Z-pyrimidinyl (1H5 H 3-p \'rida7.inyl n r. H 7-intlazolyl H S H 3-indol \l (EH1 H methyl (,,H H hutyl C,,H H 4-propylphcnyl C,;H H hydrogen C..H H 2.6-dichl0l'UPhC1LYi (7H,- H phcnyl 3BrC,;H H 3,4 dichIorophcnyI 4-ICH H 3.4-dimethoxyphenyl 2-NO2-C,;H4 H 4-fluorophcnyl 4-CH;;C;H4 H 4-hydroxyphenyl .'Z---(CH:.)- .NC.;H4 H 4-aminophcnyl 4H- ,NSO C|;H4 H 4-fluorophcnyl 4--t-C,H,,C,,H, H 4-mcthylphenyl 2,3.f (CH C,;H. H Z-nitrophcnyl n n H Z-cyanophcinyl (3H3 H 4-acctamidophcnyl (32H; CH" 3-nitrophenyl C -,H, H 2-hromophcnyl l-naphthyl H 4-bromophenyl B-thienyl H 4-cyanophcnyl Z-thicnyl H Z-methylphenyl 2furyl H 3.5-dimcthoxyphenyl 3-indolyl H 2chlorophenyl :i z)n H 3-bromophcnyl C- H C H 4-nitrophcnyl C. H -,CH CH H 3.4-dichlorophcnyl 4HOC.;H, H phenyl l ,4-cyclohexadicnyl H 3bromophcnyl (CH2) Z-cyanophcnyl (CH- 4-nitrophenyl (CH2)X 4-fluorophcnyl cycloheptatrienyl H phcnyl EXAMPLE Vl 3,4-Dichlorobenzylidene Ethyl Hydrogen Malonate 3,4-Dichlorobenzaldehyde (0.010 mole) is mixed with 0.010 mole of monoethyl malonate (Organic Syntheses Coll. Vol. IV, p. 417) and heated on the steam bath for 15 hours. The material is suspended in ether and extracted with 2 portions of sodium bicarbonate solution. The separated basic aqueous layer is overlayed with ether and acidified with 6 N HCl. The ether layer is dried over-anhydrous sodium sulfate, filtered and taken to dryness under vacuum. The white solid is crystallized from cyclohexane, yield 32%.

6-[ D-2-Phenyl-2-( Ethyl-3,4-Dichlorobenzylidenemalonamido)AcetamidolPenicillanic Acid Triethylamine 3,4Dichlorobenzylidene ethyl hydrogen malonic acid (0.003 mole) in 20 ml. of water is cooled to. 0C. and adjusted to pH 6.0 to give a clear solution. To this solution is added 0.003 mole of (u-a-aminobenzylpenicillin and 0.003 mole of lethyl-3-(dimethylaminopropyllcarbodiimide hydrochloride. The reaction mixture, maintained at pH 6.0 with 6 N HCI, is stirred at 0C for about 30 minutes, followed by stirring at room temperature for 4 hours. The solution is adjusted to pH 7.7 with saturated sodium bicarbonate solution EXAMPLE Vll Mono-5-lndanyl Malonate Malonic acid (0.108 mole) in 100 ml. of ether is refluxed for 1 hour with 0.120 mole of thionyl chloride. After cooling to room temperature, 0.108 mole of 5- indanol is added and the yellow solution is refluxed for 2 hours. The reaction solution is cooled to room trmperature and the excess thionyl chloride is removed under vacuum. Ether (50 ml.) is added and the solution is washed twice with 50 ml. portions of water. The ether layer is extracted with two 50 ml. portions of sodium bicarbonate solution and one 25 ml. portion of water. The combined basic aqueous phase is acidified with 6 N HCl and extracted with several portions of ether. The ether extract is dried over anhydrous sodium sulfate, filtered and taken to dryness under vacuum to a viscous yellow which crystallizes on standing, m.p. 51-54C., yield 54%.

3,4-Dichlorobenzylidene 5- lndanyl Hydrogen Malonate Mono-S-indanyl malonate (0.010 mole) is mixed with 0.010 mole of 3,4-dichlorobenzalydehyde and heated on a steam bath for about 18 hours. The crystals obtained after cooling the reaction mixture to room temperature are recrystallized from acetonitrile, m.p. l74178C., yield 33%.

6-[ D-2-Phenyl-2-( 5 -Indanyl-3,4-Dichlorobenzylidene malonamido)Acetamido]Penicillanic Acid Triethylamine 3,4-Dichlorobenzylidene 5-indanyl hydrogen malonate (0.0031 mole) in 30 ml. of water is adjusted to pH 6.0 to give a torbid solution. To this solution is added 0.0031 mole of b-oz-aminobenzylpenicillin and 0.0031 mole of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride. After stirring for 30 minutes at C., maintaining the pH at 6.0 with 6 N HCl, the solution is stirred for about 3 hours at room temperature. The pH is adjusted to 7.7 with saturated sodium bicarbonate solution and extracted with 2 portions of ethyl acetate. The aqueous phase is overlayed with ethyl acetate and the pH adjusted to 2.7 with 6 N HCl. The separated ethyl acetate extract is washed once with water and once with saturated sodium chloride solution. To the ethyl acetate extrat, dried over anhydrous sodium sulfate, is added 0.0031 mole of triethylamine and the solution taken to dryness under vacuum. Ethyl acetate and triethylamine are removed from the white gum by treating azeotropically with 3 portions of methylene chloride, yield 20%.

EXAMPLE V111 p-Acetamidobenzylidene 5'-Indanyl Hydrogen Malonate p-Acetamidobenzaldehyde (Eastman) is reacted with mono-5-indanyl malonate by the method of Example V to yield crystals, m.p. l194C., yield 16%.

6-[D-2-Phenyl-2-( 5 '-lndanyl-p-acetamidobenzylidenemalonamido)-AcetamidolPenicillanic Acid Triethylamine 6-a-D-aminobenzylpenicillin is reacted with pacetamidebenzylidene 5-indanyl hydrogen malonate by the method of Example VII.

Repitition of this procedure but replacing pacetamidobenzylidene 5-indanyl hydrogen malonate with equimolar amounts of the appropriate malonic acid derivative of the formula to provide a penicillin of the formula 2-chlorophcnyl 2 (1,4-naphthoquinonyl) EXAMPLE lX The method of Example V1 is repeated replacing 6-a-D-aminobenzylpenicillin with equimolar amounts of the appropriate ester of 6a-D-aminobenzylpenicillin producing compounds of the formula wherein R and Q are the same as those described in Example V111, and R is as described in Example Ill.

EXAMPLE XI The -free mono-acids are transformed to their monosodium, potassium, calcium, magnesium, ammonium,

GENERAL PROCEDURE FOR a-AMlNOACYLPENlClLLlN ACYLOXYALKYL ESTERS The procedure of Daehne et al., J. Med. Chem. 13, 612 1970) is employed to prepare the title compounds. The procedure comprises acylating the appropriate acyloxyalkyl -aminopenicillanate with the ap propriate amino acid chloride hydrochloride in a sol vent such as methylene chloride in the presence of an acid acceptor (NaHCO The ester, amino acid chloride hydrochloride and NaHCO are reacted in a molar proportion of about l.25:l.():2.5 at l0C. for 2 3 hours. The mixture is filtered through diatomaceous earth, isopropanol added to the filtrate which is then concentrated in vacuo. Concentration is continued until the produce separates. lsopropanol and ether are added to the mixture to complete precipitation of the product.

What is claimed is:

l. A compound of the formula wherein R is selected from the group consisting of hydroen and acyloxy lower alkyl wherein acyloxy is selected from the group consisting of lower alkanoyloxy, benzoyloxy and substituted benzoyloxy wherein the substituent is selected from the group consisting of chloro, bromo, fluoro, lower alkyl, lower alkoxy and trifluoromethyl, cations selected from the group consisting of sodium, potassium, calcium, magnesium and ammonium; and amines selected from the group consisting of procaine, N,N'-dibenzylethylenediamine, N-ethylpiperidine, dibenzylamine, N,N-bis(dehydroabietyl)ethylenediamine, l-ephenamine, triethylamine and benzhydrylamine;

R is selected from the group consisting of hydrogen, alkyl of from I to 14 carbon atoms, cycloalkyl of from 3 to 6 carbon atoms, 1,4-cyclohexadienyl,

18 cyclohepatrienyl, naphthyl, benzyl, phenethyl, indolylmethyl, furyl, thienyl, w-ethylthio(lower)alkyl and wherein Y is selected from the group consisting of hydrogen, nitro, di(lower alkyl) amino, lower alkanoylamino, lower alkyl, lower alkoxy, hydroxy, sulfamyl, chloro, bromo, fluoro, iodo and trifluoromethyl;

R is selected from the group consisting of hydrogen and lower alkyl;

R and R when taken together with the carbon atom to which they are attached are cycloalkylidene of 3 to 10 carbon atoms;

R is a member selected from the group consisting of hydrogen, alkyl having from 1 to 4 carbon atoms and cycloalkyl having from 3 to 6 carbon atoms; and

Q is a member selected from the group consisting of hydrogen, alkyl having from 1 to 4 carbon atoms, naphthyl, quinolyl, pphenyl, biphenyl, indanyl, 2-( 1,4-naphthoquinonyl); cations selected from the grooup consisting of sodium, potassium, calcium, magnesium and ammonium; and amines selected from the group consisting of procaine, N,N- dibenzylethylenediamine, N-ethylpiperidine, dibenzylamine, N,N-bis(dehydroabietyl)ethylenediamine, l-ephenamine, triethylamine and benzhydrylamine.

2. A compound of the formula O=C N CH-COOR (i-COOQ C malonamido)acetamido]penicillanic acid. 

1. A COMPOUND OF THE FORMULA
 2. A compound of the formula
 3. 6-(D-2-Phenyl-2-(2''-thienylmalonamido)acetamido)penicillanic acid. 